Multiplex immunoassay for serological diagnosis of Mycobacterium bovis infection in cattle.
Efforts to develop a better diagnostic assay for bovine tuberculosis have shown that the sensitivity and specificity of an assay can be improved by the use of two or more antigens. As reported here, we developed a multiplex chemiluminescent immunoassay that can simultaneously detect antibody activity to 25 antigens in a single well in a 96-well plate array format. The chemiluminescent signal is captured with a digital imaging system and analyzed with a macro program that tracks each serum for its pattern of antibody activity for Mycobacterium bovis antigens. The comparison of sera from 522 infected and 1,489 uninfected animals showed that a sensitivity of 93.1% and a specificity of 98.4% can be achieved with a combination of antigens. The assay system is rapid and can be automated for use in a centralized laboratory.
Whelan
Clare
C
Enfer Scientific, Unit T, M7 Business Park, Newhall, Naas, County Kildare, Ireland.
Shuralev
Eduard
E
O'Keeffe
Grainne
G
Hyland
Paula
P
Kwok
Hang Fai
HF
Snoddy
Philip
P
O'Brien
Amanda
A
Connolly
Marie
M
Quinn
Padraig
P
Groll
Matt
M
Watterson
Todd
T
Call
Sara
S
Kenny
Kevin
K
Duignan
Anthony
A
Hamilton
Mary Jo
MJ
Buddle
Bryce M
BM
Johnston
James A
JA
Davis
William C
WC
Olwill
Shane A
SA
Clarke
John
J
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D002417:Cattle; D007118:Immunoassay; D008163:Luminescent Measurements / Q000379:methods*; D009163:Mycobacterium bovis / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014380:Tuberculosis, Bovine / Q000175:diagnosis*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins